Retana Carmen, Sanchez Elsa I, Gonzalez Sirenia, Perez-Lopez Alejandro, Cruz Armando, Lagunas-Munoz Jesus, Alfaro-Cruz Carmen, Vital-Flores Socorro, Reyes José L
Pharmacology Department Centre for Research and Advanced Studies National Polytechnic Institute, Mexico, D.F., Mexico.
PLoS One. 2013 Nov 4;8(11):e79678. doi: 10.1371/journal.pone.0079678. eCollection 2013.
Patients undergoing continuous ambulatory peritoneal dialysis are classified according to their peritoneal permeability as low transporter (low solute permeability) or High transporter (high solute permeability). Factors that determine the differences in permeability between them have not been fully disclosed. We investigated morphological features of cultured human peritoneal mesothelial cells from low or high transporter patients and its response to All trans retinoic Acid (ATRA, vitamin A active metabolite), as compared to non-uremic human peritoneal mesothelial cells. Control cells were isolated from human omentum. High or low transporter cells were obtained from dialysis effluents. Cells were cultured in media containing ATRA (0, 50, 100 or 200 nM). We studied length and distribution of microvilli and cilia (scanning electron microscopy), epithelial (cytokeratin, claudin-1, ZO-1 and occludin) and mesenchymal (vimentin and α-smooth muscle actin) transition markers by immunofluorescence and Western blot, and transforming growth factor β1 expression by Western blot. Low and high transporter exhibited hypertrophic cells, reduction in claudin-1, occludin and ZO-1 expression, cytokeratin and vimentin disorganization and positive α-smooth muscle actin label. Vimentin, α-smooth muscle actin and transforming growth factor-β1 were overexpressed in low transporter. Ciliated cells were diminished in low and high transporters. Microvilli number and length were severely reduced in high transporter. ATRA reduced hypertrophic cells number in low transporter. It also improved cytokeratin and vimentin organization, decreased vimentin and α-smooth muscle actin expression, and increased claudin 1, occludin and ZO-1 expression, in low and high transporter. In low transporter, ATRA reduced transforming growth factor-β1 expression. ATRA augmented percentage of ciliated cells in low and high transporter. It also augmented cilia length in high transporter. Alterations in structure, epithelial mesenchymal markers and transforming growth factor-β1 expression were differential between low and high transporter. Beneficial effects of ATRA were improved human peritoneal mesothelial cells morphology tending to normalize structures.
接受持续性非卧床腹膜透析的患者根据其腹膜通透性被分为低转运者（低溶质通透性）或高转运者（高溶质通透性）。决定两者通透性差异的因素尚未完全明了。我们研究了来自低转运者或高转运者患者的培养人腹膜间皮细胞的形态特征及其对全反式维甲酸（ATRA，维生素A的活性代谢产物）的反应，并与非尿毒症人腹膜间皮细胞进行比较。对照细胞从人网膜中分离获得。高转运者或低转运者细胞从透析流出液中获取。细胞在含有ATRA（0、50、100或200 nM）的培养基中培养。我们通过扫描电子显微镜研究微绒毛和纤毛的长度及分布，通过免疫荧光和蛋白质印迹法研究上皮（细胞角蛋白、闭合蛋白-1、紧密连接蛋白-1和闭合蛋白）和间充质（波形蛋白和α-平滑肌肌动蛋白）转化标志物，并通过蛋白质印迹法研究转化生长因子β1的表达。低转运者和高转运者均表现出细胞肥大、闭合蛋白-1、闭合蛋白和紧密连接蛋白-1表达减少、细胞角蛋白和波形蛋白紊乱以及α-平滑肌肌动蛋白标记阳性。波形蛋白、α-平滑肌肌动蛋白和转化生长因子-β1在低转运者中过表达。低转运者和高转运者中的纤毛细胞减少。高转运者中的微绒毛数量和长度严重减少。ATRA减少了低转运者中肥大细胞的数量。它还改善了低转运者和高转运者中细胞角蛋白和波形蛋白的排列，降低了波形蛋白和α-平滑肌肌动蛋白的表达，并增加了闭合蛋白1、闭合蛋白和紧密连接蛋白-1的表达。在低转运者中，ATRA降低了转化生长因子-β1的表达。ATRA增加了低转运者和高转运者中纤毛细胞的百分比。它还增加了高转运者中纤毛的长度。低转运者和高转运者在结构、上皮间充质标志物和转化生长因子-β1表达方面的改变存在差异。ATRA的有益作用是改善了人腹膜间皮细胞的形态，使其结构趋于正常化。
